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Histone H3 monomethyl Lys4 antibody (mAb)
| Clone: MABI 0302 |
| Catalog No: 39635 | Format: 100 µg | $340 | Buy Now |
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Applications Table
| Western Blot | Chromatin IP |
|---|---|
| 0.5 - 2 µg/ml | 5 - 10 µg per ChIP |
for Histone H3 monomethyl Lys4 antibody (mAb) (Clone MABI 0302)Background
Histone H3 is one of the core components of the nucleosome. The nucleosome is the smallest subunit of chromatin and consists of 147 base pairs of DNA wrapped around an octamer of core histone proteins (two each of Histone H2A, Histone H2B, Histone H3 and Histone H4). Histone H1 is a linker histone, present at the interface between the nucleosome core and DNA entry/exit points. Histone H1 is responsible for establishing higher-order chromatin structure.
Chromatin is subject to a variety of chemical modifications, including post-translational modifications of the histone proteins and the methylation of cytosine residues in the DNA. Reported histone modifications include acetylation, methylation, phosphorylation, ubiquitylation, glycosylation, ADP-ribosylation, carbonylation and SUMOylation; these modifications play a major role in regulating gene expression.
The methylation of histones can occur on two different residues: arginine or lysine. Histone methylation can be associated with transcriptional activation or repression, depending on the methylated residue. Lysine 4 of histone H3 can be mono-, di- or trimethylated by different histone methyltransferases (HMTs) such as SET1 or ASH1. Methylation of Lys4 is often associated with transcriptional activation. The demethylase LSD1 is able to demethylate histone H3 Lys4.
for Histone H3 monomethyl Lys4 antibody (mAb) (Clone MABI 0302)Immunogen
The Histone H3 monomethyl Lys4 antibody was raised against a peptide containing monomethyl lysine 4 of human histone H3.
for Histone H3 monomethyl Lys4 antibody (mAb) (Clone MABI 0302)Application Notes
This Histone H3 monomethyl Lys4 antibody (H3K4me) has been validated for use in Western blotting (0.5 - 2 µg/ml) and chromatin IP (5 - 10 µg per ChIP).
for Histone H3 monomethyl Lys4 antibody (mAb) (Clone MABI 0302)Buffer
PBS pH 7.5, 30% glycerol, 0.035% sodium azide.
for Histone H3 monomethyl Lys4 antibody (mAb) (Clone MABI 0302)Positive Control
Active Motif's HeLa acid extract (Catalog No. 36200) can be used as a positive control for Histone H3 monomethyl Lys4 antibody (Clone MABI 0302).
Chromatin IP Assays
Active Motif has a variety of Chromatin Immunoprecipitation Kits for every need.
Please visit our Chromatin IP Page to learn about ChIP products designed for use with Histone H3 monomethyl Lys4 antibody (mAb).
Bridging Antibody to Improve ChIP, MeDIP and IP
Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use Histone H3 monomethyl Lys4 antibody (mAb).
for Histone H3 monomethyl Lys4 antibody (mAb) (Clone MABI 0302)References
This antibody was used in ChIP assays as described in Kimura, H. et al (2008) Cell Struct Func 33: 61-73.
Histone H3 monomethyl Lys4 antibody (mAb) tested by Western blot.
HeLa acid extract probed with Histone H3 monomethyl Lys4 antibody (mAb) (2 µg/ml dilution).
Histone H3 monomethyl Lys4 antibody (mAb) tested by ChIP.
Chromatin IP performed using the ChiP-IT Express Kit (Catalog No. 53008) and 100 µl of Ready-to-ChIP HeLa Chromatin (Catalog No. 53015) per ChIP. Subsequent to the ChIP reaction, DNA was purified from the immunoprecipitated chromatin and a region of the human GAPDH promoter was amplified by PCR.
- Lane 1: ChIP using 10 µg of Histone H3 monomethyl Lys4 antibody (mAb).
- Lane 2: ChIP using negative control mouse IgG.
- Lane 3: PCR input control.
Histone H3 monomethyl Lys4 antibody (mAb) tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 monomethyl Lys4 antibody (mAb) for monomethyl-Lys4 histone H3. Methylated peptides corresponding to the immunogen and related peptides were spotted onto PVDF and probed with the antibody at 1 µg/ml. The amount of peptide (picomoles) spotted is indicated next to each row.
Top panel:
Lane 1: Unmodified lysine 4 peptide.
Lane 2: Monomethyl lysine 4 peptide.
Lane 3: Dimethyl lysine 4 peptide.
Lane 4: Trimethyl lysine 4 peptide.
Lane 5: Unmodified peptide corresponding to amino acids 6-22 of human histone H3.
Lane 6: Monomethyl lysine 9 peptide.
Lane 7: Dimethyl lysine 9 peptide.
Lane 8: Trimethyl lysine 9 peptide.
Bottom panel:
Lane 1: dimethyl lysine 14 peptide.
Lane 2: monomethyl lysine 18 peptide.
Lane 3: dimethyl lysine 18 peptide.
Lane 4: trimethyl lysine 18 peptide.
Lane 5: unmodified peptide corresponding to amino acids 18-27 of human histone H3.
Lane 6: monomethyl lysine 23 peptide.
Lane 7: dimethyl lysine 23 peptide.
Lane 8: trimethyl lysine 23 peptide.
Lane 9: unmodified peptide corresponding to amino acids 22-32 of human histone H3.
Lane 10: monomethyl lysine 27 peptide.
Lane 11: dimethyl lysine 27 peptide.
Lane 12: trimethyl lysine 27 peptide.
for Histone H3 monomethyl Lys4 antibody (mAb) (Clone MABI 0302)Storage
Antibodies in solution can be stored at -20°C for 2 years. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
for Histone H3 monomethyl Lys4 antibody (mAb) (Clone MABI 0302)Guarantee
This product is guaranteed for 6 months from date of receipt.
This product is for research use only and is not for use in diagnostic procedures.
Application Key
- ChIP = Chromatin Immunoprecipitation;
- DB = Dot Blot;
- ELISA = Enzyme-linked Immunosorbent Assay;
- EMSA = Electrophoretic Mobility Shift Assay
- FACS = Flow Cytometry;
- ICC = Immunocytochemistry;
- IF = Immunofluorescence;
- IHC = Immunohistochemistry;
- IP = Immunoprecipitation;
- MeDIP = Methyl-DNA Immunoprecipitation;
- WB = Western Blotting
